Lettuce line RX 16883035

ABSTRACT

The invention provides seed and plants of the lettuce line designated RX 16883035. The invention thus relates to the plants, seeds and tissue cultures of lettuce line RX 16883035, and to methods for producing a lettuce plant produced by crossing a plant of lettuce line RX 16883035 with itself or with another lettuce plant, such as a plant of another line. The invention further relates to seeds and plants produced by such crossing. The invention further relates to parts of a plant of lettuce line RX 16883035, including the gametes of such plants.

FIELD OF THE INVENTION

The present invention relates to the field of plant breeding and, morespecifically, to the development of lettuce line RX 16883035.

BACKGROUND OF THE INVENTION

The goal of vegetable breeding is to combine various desirable traits ina single variety/hybrid. Such desirable traits may include greateryield, resistance to insects or pests, tolerance to heat and drought,better agronomic quality, higher nutritional value, growth rate andfruit properties.

Breeding techniques take advantage of a plant's method of pollination.There are two general methods of pollination: a plant self-pollinates ifpollen from one flower is transferred to the same or another flower ofthe same plant or plant variety. A plant cross-pollinates if pollencomes to it from a flower of a different plant variety.

Plants that have been self-pollinated and selected for type over manygenerations become homozygous at almost all gene loci and produce auniform population of true breeding progeny, a homozygous plant. A crossbetween two such homozygous plants of different varieties produces auniform population of hybrid plants that are heterozygous for many geneloci. Conversely, a cross of two plants each heterozygous at a number ofloci produces a population of hybrid plants that differ genetically andare not uniform. The resulting non-uniformity makes performanceunpredictable.

The development of uniform varieties requires the development ofhomozygous inbred plants, the crossing of these inbred plants, and theevaluation of the crosses. Pedigree breeding and recurrent selection areexamples of breeding methods that have been used to develop inbredplants from breeding populations. Those breeding methods combine thegenetic backgrounds from two or more plants or various other broad-basedsources into breeding pools from which new lines are developed byselfing and selection of desired phenotypes. The new lines are evaluatedto determine which of those have commercial potential.

SUMMARY OF THE INVENTION

In one aspect, the present invention provides a lettuce plant of theline designated RX 16883035. Also provided are lettuce plants having thephysiological and morphological characteristics of the lettuce linedesignated RX 16883035. Parts of the lettuce plant of the presentinvention are also provided, for example, including pollen, an ovule,and a cell of the plant.

The invention also concerns seed of lettuce line RX 16883035. Thelettuce seed of the invention may be provided as an essentiallyhomogeneous population of lettuce seed of the line designated RX16883035. Essentially homogeneous populations of seed are generally freefrom substantial numbers of other seed. In certain embodiments of theinvention, seed of line RX 16883035 may be provided forming at leastabout 97% of the total seed, including at least about 98%, 99%, or moreof the seed. The population of lettuce seed may be particularly definedas being essentially free from hybrid seed. The seed population may beseparately grown to provide an essentially homogeneous population oflettuce plants designated RX 16883035.

In another aspect of the invention, a plant of lettuce line RX 16883035comprising an added heritable trait is provided. The heritable trait maycomprise a genetic locus that is a dominant or recessive allele. In oneembodiment of the invention, a plant of lettuce line RX 16883035 isdefined as comprising a single locus conversion. In specific embodimentsof the invention, an added genetic locus confers one or more traits suchas, for example, herbicide tolerance, insect resistance, diseaseresistance, and modified carbohydrate metabolism. The trait may be, forexample, conferred by a naturally occurring gene introduced into thegenome of the line by backcrossing, a natural or induced mutation, or atransgene introduced through genetic transformation techniques into theplant or a progenitor of any previous generation thereof. Whenintroduced through transformation, a genetic locus may comprise one ormore transgenes integrated at a single chromosomal location.

In another aspect of the invention, a tissue culture of regenerablecells of a plant of line RX 16883035 is provided. The tissue culturewill preferably be capable of regenerating plants capable of expressingall of the physiological and morphological characteristics of the line,and of regenerating plants having substantially the same genotype asother plants of the line. Examples of some of the physiological andmorphological characteristics of the line RX 16883035 include thosetraits set forth in the tables herein. The regenerable cells in suchtissue cultures may be derived, for example, from embryos, meristems,cotyledons, pollen, leaves, anthers, roots, root tips, pistil, flower,seed and stalks. Still further, the present invention provides lettuceplants regenerated from a tissue culture of the invention, the plantshaving all the physiological and morphological characteristics of lineRX 16883035.

In yet another aspect of the invention, processes are provided forproducing lettuce seeds and plants, which processes generally comprisecrossing a first parent lettuce plant with a second parent lettuceplant, wherein at least one of the first or second parent lettuce plantsis a plant of the line designated RX 16883035. These processes may befurther exemplified as processes for preparing hybrid lettuce seed orplants, wherein a first lettuce plant is crossed with a second lettuceplant of a different, distinct line to provide a hybrid that has, as oneof its parents, the lettuce plant line RX 16883035. In these processes,crossing will result in the production of seed. The seed productionoccurs regardless of whether the seed is collected or not.

In one embodiment of the invention, the first step in “crossing”comprises planting seeds of a first and second parent lettuce plant,often in proximity so that pollination will occur for example, mediatedby insect vectors. Alternatively, pollen can be transferred manually.Where the plant is self-pollinated, pollination may occur without theneed for direct human intervention other than plant cultivation.

A second step may comprise cultivating or growing the seeds of first andsecond parent lettuce plants into plants that bear flowers. A third stepmay comprise preventing self-pollination of the plants, such as byemasculating the male portions of flowers, (i.e., treating ormanipulating the flowers to produce an emasculated parent lettuceplant). Self-incompatibility systems may also be used in some hybridcrops for the same purpose. Self-incompatible plants still shed viablepollen and can pollinate plants of other varieties but are incapable ofpollinating themselves or other plants of the same line.

A fourth step for a hybrid cross may comprise cross-pollination betweenthe first and second parent lettuce plants. Yet another step comprisesharvesting the seeds from at least one of the parent lettuce plants. Theharvested seed can be grown to produce a lettuce plant or hybrid lettuceplant.

The present invention also provides the lettuce seeds and plantsproduced by a process that comprises crossing a first parent lettuceplant with a second parent lettuce plant, wherein at least one of thefirst or second parent lettuce plants is a plant of the line designatedRX 16883035. In one embodiment of the invention, lettuce seed and plantsproduced by the process are first generation (F₁) hybrid lettuce seedand plants produced by crossing a plant in accordance with the inventionwith another, distinct plant. The present invention further contemplatesplant parts of such an F₁ hybrid lettuce plant, and methods of usethereof. Therefore, certain exemplary embodiments of the inventionprovide an F₁ hybrid lettuce plant and seed thereof.

In still yet another aspect of the invention, the genetic complement ofthe lettuce plant line designated RX 16883035 is provided. The phrase“genetic complement” is used to refer to the aggregate of nucleotidesequences, the expression of which sequences defines the phenotype of,in the present case, a lettuce plant, or a cell or tissue of that plant.A genetic complement thus represents the genetic makeup of a cell,tissue or plant, and a hybrid genetic complement represents the geneticmake up of a hybrid cell, tissue or plant. The invention thus provideslettuce plant cells that have a genetic complement in accordance withthe lettuce plant cells disclosed herein, and plants, seeds and plantscontaining such cells.

Plant genetic complements may be assessed by genetic marker profiles,and by the expression of phenotypic traits that are characteristic ofthe expression of the genetic complement, e.g., isozyme typing profiles.It is understood that line RX 16883035 or a first generation progenythereof could be identified by any of the many well known techniquessuch as, for example, Simple Sequence Length Polymorphisms (SSLPs)(Williams et al., Nucleic Acids Res., 1 8:6531-6535, 1990), RandomlyAmplified Polymorphic DNAs (RAPDs), DNA Amplification Fingerprinting(DAF), Sequence Characterized Amplified Regions (SCARs), ArbitraryPrimed Polymerase Chain Reaction (AP-PCR), Amplified Fragment LengthPolymorphisms (AFLPs) (EP 534 858, specifically incorporated herein byreference in its entirety), and Single Nucleotide Polymorphisms (SNPs)(Wang et al., Science, 280:1077-1082, 1998).

In still yet another aspect, the present invention provides hybridgenetic complements, as represented by lettuce plant cells, tissues,plants, and seeds, formed by the combination of a haploid geneticcomplement of a lettuce plant of the invention with a haploid geneticcomplement of a second lettuce plant, preferably, another, distinctlettuce plant. In another aspect, the present invention provides alettuce plant regenerated from a tissue culture that comprises a hybridgenetic complement of this invention.

In still yet another aspect, the invention provides a method ofdetermining the genotype of a plant of lettuce line RX 16883035comprising detecting in the genome of the plant at least a firstpolymorphism. The method may, in certain embodiments, comprise detectinga plurality of polymorphisms in the genome of the plant. The method mayfurther comprise storing the results of the step of detecting theplurality of polymorphisms on a computer readable medium. The inventionfurther provides a computer readable medium produced by such a method.

In still yet another aspect, the present invention provides a method ofproducing a plant derived from line RX 16883035, the method comprisingthe steps of: (a) preparing a progeny plant derived from line RX16883035, wherein said preparing comprises crossing a plant of the lineRX 16883035 with a second plant; and (b) crossing the progeny plant withitself or a second plant to produce a seed of a progeny plant of asubsequent generation. In further embodiments, the method mayadditionally comprise: (c) growing a progeny plant of a subsequentgeneration from said seed of a progeny plant of a subsequent generationand crossing the progeny plant of a subsequent generation with itself ora second plant; and repeating the steps for an additional 3-10generations to produce a plant derived from line RX 16883035. The plantderived from line RX 16883035 may be an inbred line, and theaforementioned repeated crossing steps may be defined as comprisingsufficient inbreeding to produce the inbred line. In the method, it maybe desirable to select particular plants resulting from step (c) forcontinued crossing according to steps (b) and (c). By selecting plantshaving one or more desirable traits, a plant derived from line RX16883035 is obtained which possesses some of the desirable traits of theline as well as potentially other selected traits.

In certain embodiments, the present invention provides a method ofproducing lettuce comprising: (a) obtaining a plant of lettuce line RX16883035, wherein the plant has been cultivated to maturity, and (b)collecting lettuce from the plant.

Any embodiment discussed herein with respect to one aspect of theinvention applies to other aspects of the invention as well, unlessspecifically noted.

The term “about” is used to indicate that a value includes the standarddeviation of error for the device or method being employed to determinethe value. The use of the term “or” in the claims is used to mean“and/or” unless explicitly indicated to refer to alternatives only orthe alternatives are mutually exclusive, although the disclosuresupports a definition that refers to only alternatives and to “and/or.”When used in conjunction with the word “comprising” or other openlanguage in the claims, the words “a” and “an” denote “one or more,”unless specifically noted. The terms “comprise,” “have” and “include”are open-ended linking verbs. Any forms or tenses of one or more ofthese verbs, such as “comprises,” “comprising,” “has,” “having,”“includes” and “including,” are also open-ended. For example, any methodthat “comprises,” “has” or “includes” one or more steps is not limitedto possessing only those one or more steps and also covers otherunlisted steps. Similarly, any plant that “comprises,” “has” or“includes” one or more traits is not limited to possessing only thoseone or more traits and covers other unlisted traits.

Other objects, features and advantages of the present invention willbecome apparent from the following detailed description. It should beunderstood, however, that the detailed description and any specificexamples provided, while indicating specific embodiments of theinvention, are given by way of illustration only, since various changesand modifications within the spirit and scope of the invention willbecome apparent to those skilled in the art from this detaileddescription.

DETAILED DESCRIPTION OF THE INVENTION

The invention provides methods and compositions relating to plants,seeds and derivatives of lettuce line RX 16883035. This line showsuniformity and stability within the limits of environmental influencefor the traits described hereinafter. Lettuce line RX 16883035 providessufficient seed yield. By crossing with a distinct second plant, uniformF1 hybrid progeny can be obtained.

Lettuce (Lactuca sativa L.) variety RX 16883035 is resistant to DownyMildew (Bremia lactucae) race B1:1 up to and including B1:23. RX16883035 is also resistant to race B1:25.

A. Origin and Breeding History of Lettuce Line RX 16883035

Lettuce cultivar RX16883035 was developed at the SEMINIS researchstation in NIMES, France. The initial cross was made in Year 1 betweenFrillice, a commercial cutting lettuce variety from Seminis as femaleparent, and a breeding line with resistance to B1 1-23 and 25 as maleparent.

The resulting F2 plants were selected for plant type and Bremiaresistance. These plants were then self-pollinated for 2 generations toform F4 lines with selection for Bremia resistance and plant type. Atthe F4 generation the line was fixed for Bremia resistance. The F4 linewas self-pollinated three more times with selection for plant type toproduce F7 lines. The F7 line 20051486 was selected as the most uniformline, and was used as a source for seed increase for lettuce varietyRX16883035 in Year 10.

B. Physiological and Morphological Characteristics of Lettuce Line RX16883035

In accordance with one aspect of the present invention, there isprovided a plant having the physiological and morphologicalcharacteristics of lettuce line RX 16883035. A description of thephysiological and morphological characteristics of lettuce line RX16883035 is presented in Table 1.

TABLE 1 Physiological and Morphological Characteristics of Line RX16883035 Comparison Variety #1- Comparison Waldmann's Variety #2-CHARACTERISTIC RX 16883035 Green Frillice 1. Type Cutting/LeafCutting/Leaf Cutting/Leaf 2. Seed color white (US = Silver black (US:Grey white (US = Silver Gray) [TG = Verpia] Brown) [TG: Gray) Kagraner[TG = Verpia] Sommer] light dormancy light not required light notrequired light not required heat dormancy susceptible susceptiblesusceptible 3. Seedling anthocyanin coloration absent (Verpia) absent(Verpia) absent (Verpia) size of cotyledon (fully medium (Expresse)large (Verpia) medium developed) (Expresse) shape of cotyledon narrowelliptic broad elliptic narrow elliptic (Calmar) (Fiorella, (Calmar)Sunrise) 4. Leaf shape of cotyledons spatulate broad spatulate shape offourth leaf pinnately oval pinnately length/width index of 9.6 + 21.111.8 10.3 + 21 fourth leaf (length/ width × 10) apical margin (cotyledoncoarsely dentate finely dentate coarsely dentate to 4^(th) leaf stage)basal margin (cotyledon coarsely dentate moderately coarsely dentate to4^(th) leaf stage) dentate undulation (cotyledon to medium medium medium4^(th) leaf stage green color (cotyledon dark green medium green darkgreen to 4^(th) leaf stage) anthocyanin distribution absent absentabsent (cotyledon to 4^(th) leaf stage) rolling (cotyledon to 4^(th)absent absent absent leaf stage) cupping (cotyledon to slight uncuppedslight 4^(th) leaf stage) reflexing (cotyledon to apical margin apicalmargin apical margin 4^(th) leaf stage) attitude at 10-12 leafsemi-erect (Great semi-erect (Great semi-erect (Great stage Lakes 118,Soraya) Lakes 118, Lakes 118, Soraya) Soraya) leaf blade: division atentire (Fiorella, entire (Fiorella, entire (Fiorella, 10-12 leaf stageSunrise) Sunrise) Sunrise) leaf blade: incisions of present (Calmar,present (Calmar, present (Calmar, margin on apical part Gloire duDauphiné, Gloire du Gloire du Unicum) Dauphiné, Dauphiné, Unicum)Unicum) leaf blade: depth of moderate/medium absent/shallow deep (US:Great incisions on margin on (US: Vanguard) (US: Dark Green Lakes 659)[TG: apical part (harvest (TG: Ithaca Great Boston) [TG: Lagon, Monet]mature outer leaves) Lakes) Pentared, Unicum] leaf blade: density ofdense (Grand sparse (Maravilla dense (Grand incisions on margin onRapids, Ithaca Great de Verano) Rapids, Ithaca apical part Lakes) GreatLakes) leaf blade: venation flabellate (Gloire du flabellate (Gloireflabellate (Gloire Dauphiné, Locarno, du Dauphiné, du Dauphiné, Monet)Locarno, Monet) Locarno, Monet) mature leaves: deeply dentate shallowlydentate deeply dentate indentation (finest (Great Lakes 659) (GreatLakes 65) (Great Lakes divisions of the margin) 659) (harvest matureouter leaves) green color (harvest dark green medium green dark greenmature outer leaves) (Vanguard) (Great Lakes) (Vanguard) hue of greencolor of absent (Donatello, yellowish (Dorée absent outer leaves Verpia)de printemps) (Donatello, Verpia) intensity of color of dark medium darkouter leaves anthocyanin coloration absent [TG: Fiorella, absent [TG:absent [TG: Sunrise] Fiorella, Sunrise] Fiorella, Sunrise] size (harvestmature small medium small outer leaves) blistering (harvestabsent/slight or moderate/ absent/slight or mature outer leaves) veryweak (US: medium (US: very weak (US: Salinas) [TG: Donia, Vanguard) [TG:Salinas) [TG: Frillblond] Commodore] Donia, Frillblond] size of blisterssmall (Dorée de medium (Dustin, printemps) Sunrise) thickness (harvestthick (Frisée de medium (Dustin, thick (Frisée de mature outer leaves)Beauregard) Sunrise) Beauregard) trichomes (harvest absent (smooth)absent (smooth) absent (smooth) mature outer leaves) attitude at harvesterect (Feria, Riva) semi-erect erect (Feria, maturity (outer leaves(Amelia, Riva) from head lettuce or Toronto) adult leaves from cuttingand stem lettuce) shape transverse narrow transverse broad transversenarrow elliptic (Elvira, elliptic elliptic (Elvira, Madison) (Commodore,Madison) Fiorella) shape of tip rounded (Blonde rounded (Blonde rounded(Blonde Maraîchère, Maraîchère, Maraîchère, Maserati) Maserati)Maserati) 5. Plant spread of frame leaves   30 cm   34 cm   24 cmdiameter small (Bastion, small (Bastion, small (Bastion, Gotte à graineGotte à graine Gotte à graine blanche) blanche) blanche) height(flowering plant) tall (Danilla, Hilde II) fasciation (at floweringabsent (Calmar, present (Gotte absent (Calmar, stage) Romance) jauned'or) Romance) head formation no head (Blonde à no head (Blonde no head(Blonde couper améliorée, à couper à couper Lollo rossa) améliorée,Lollo améliorée, Lollo rossa) rossa) head shape non-heading non-headingnon-heading head weight  362 gms  390 gms  301 gms 6. Butt shape roundedrounded rounded midrib moderately raised prominently moderately raisedraised (Great Lakes 659) 7. Core diameter at base of head 22.7 mm  27.8mm  23.4 mm core height from base of 28.5 mm  41.7 mm  24.1 mm head toapex 8. Bolting first water date Aug. 3, 2011 Aug. 3, 2011 Aug. 3, 2011number of days from 133 106 145 first water date to seed stalk emergence(summer conditions) bolting class very slow rapid very slow time ofbeginning of very late (Erica, very late (Erica, bolting under long dayKinemontepas, Rex) Kinemontepas, conditions Rex) height of mature seed 125 cm  117 cm  118 cm stalk spread of bolter plant (at   40 cm   68 cm  28 cm widest point) bolter leaves straight curved straight margindentate dentate dentate color medium green dark green medium greenbolter habit: terminal present present present inflorescence bolterhabit: lateral absent present absent shoots axillary sprouting absent orvery weak strong (Riva) absent or very (Valmaine) weak (Valmaine) 9.Maturity: earliness of harvest- summer 93 days summer 92 days summer 93days mature head formation *These are typical values. Values may varydue to environment. Other values that are substantially equivalent arealso within the scope of the invention.

C. Breeding Lettuce Line RX 16883035

One aspect of the current invention concerns methods for crossing thelettuce line RX 16883035 with itself or a second plant and the seeds andplants produced by such methods. These methods can be used forpropagation of line RX 16883035, or can be used to produce hybridlettuce seeds and the plants grown therefrom. Hybrid seeds are producedby crossing line RX 16883035 with second lettuce parent line.

The development of new varieties using one or more starting varieties iswell known in the art. In accordance with the invention, novel varietiesmay be created by crossing line RX 16883035 followed by multiplegenerations of breeding according to such well known methods. Newvarieties may be created by crossing with any second plant. In selectingsuch a second plant to cross for the purpose of developing novel lines,it may be desired to choose those plants which either themselves exhibitone or more selected desirable characteristics or which exhibit thedesired characteristic(s) when in hybrid combination. Once initialcrosses have been made, inbreeding and selection take place to producenew varieties. For development of a uniform line, often five or moregenerations of selfing and selection are involved.

Uniform lines of new varieties may also be developed by way ofdouble-haploids. This technique allows the creation of true breedinglines without the need for multiple generations of selfing andselection. In this manner true breeding lines can be produced in aslittle as one generation. Haploid embryos may be produced frommicrospores, pollen, anther cultures, or ovary cultures. The haploidembryos may then be doubled autonomously, or by chemical treatments(e.g. colchicine treatment). Alternatively, haploid embryos may be growninto haploid plants and treated to induce chromosome doubling. In eithercase, fertile homozygous plants are obtained. In accordance with theinvention, any of such techniques may be used in connection with line RX16883035 and progeny thereof to achieve a homozygous line.

New varieties may be created, for example, by crossing line RX 16883035with any second plant and selection of progeny in various generationsand/or by doubled haploid technology. In choosing a second plant tocross for the purpose of developing novel lines, it may be desired tochoose those plants which either themselves exhibit one or more selecteddesirable characteristics or which exhibit the desired characteristic(s)in progeny. After one or more lines are crossed, true-breeding lines maybe developed.

Backcrossing can also be used to improve an inbred plant. Backcrossingtransfers a specific desirable trait from one inbred or non-inbredsource to an inbred that lacks that trait. This can be accomplished, forexample, by first crossing a superior inbred (A) (recurrent parent) to adonor inbred (non-recurrent parent), which carries the appropriate locusor loci for the trait in question. The progeny of this cross are thenmated back to the superior recurrent parent (A) followed by selection inthe resultant progeny for the desired trait to be transferred from thenon-recurrent parent. After five or more backcross generations withselection for the desired trait, the progeny are heterozygous for locicontrolling the characteristic being transferred, but are like thesuperior parent for most or almost all other loci. The last backcrossgeneration would be selfed to give pure breeding progeny for the traitbeing transferred.

The line of the present invention is particularly well suited for thedevelopment of new lines based on the elite nature of the geneticbackground of the line. In selecting a second plant to cross with RX16883035 for the purpose of developing novel lettuce lines, it willtypically be preferred to choose those plants which either themselvesexhibit one or more selected desirable characteristics or which exhibitthe desired characteristic(s) when in hybrid combination.

D. Further Embodiments of the Invention

In specific embodiments, the invention provides plants modified toinclude at least a first desired heritable trait. Such plants may, inone embodiment, be developed by a plant breeding technique calledbackcrossing, wherein essentially all of the desired morphological andphysiological characteristics of a variety are recovered in addition toa genetic locus transferred into the plant via the backcrossingtechnique. The terms converted plant or single locus converted plant asused herein refers to those lettuce plants which are developed by aplant breeding technique called backcrossing, wherein essentially all ofthe desired morphological and physiological characteristics of a varietyare recovered in addition to the single locus transferred into thevariety via the backcrossing technique. By essentially all of themorphological and physiological characteristics, it is meant that thecharacteristics of a plant are recovered that are otherwise present whencompared in the same environment, other than an occasional variant traitthat might arise during backcrossing or direct introduction of atransgene. It is understood that a locus introduced by backcrossing mayor may not be transgenic in origin, and thus the term backcrossingspecifically includes backcrossing to introduce loci that were createdby genetic transformation.

Backcrossing methods can be used with the present invention to improveor introduce a characteristic into the present variety. The parentallettuce plant which contributes the locus for the desired characteristicis termed the nonrecurrent or donor parent. This terminology refers tothe fact that the nonrecurrent parent is used one time in the backcrossprotocol and therefore does not recur. The parental lettuce plant towhich the locus or loci from the nonrecurrent parent are transferred isknown as the recurrent parent as it is used for several rounds in thebackcrossing protocol.

In a typical backcross protocol, the original variety of interest(recurrent parent) is crossed to a second variety (nonrecurrent parent)that carries the single locus of interest to be transferred. Theresulting progeny from this cross are then crossed again to therecurrent parent and the process is repeated until a lettuce plant isobtained wherein essentially all of the desired morphological andphysiological characteristics of the recurrent parent are recovered inthe converted plant, in addition to the single transferred locus fromthe nonrecurrent parent.

The selection of a suitable recurrent parent is an important step for asuccessful backcrossing procedure. The goal of a backcross protocol isto alter or substitute a single trait or characteristic in the originalvariety. To accomplish this, a single locus of the recurrent variety ismodified or substituted with the desired locus from the nonrecurrentparent, while retaining essentially all of the rest of the desiredgenetic, and therefore the desired physiological and morphologicalconstitution of the original variety. The choice of the particularnonrecurrent parent will depend on the purpose of the backcross; one ofthe major purposes is to add some commercially desirable trait to theplant. The exact backcrossing protocol will depend on the characteristicor trait being altered to determine an appropriate testing protocol.Although backcrossing methods are simplified when the characteristicbeing transferred is a dominant allele, a recessive allele may also betransferred. In this instance it may be necessary to introduce a test ofthe progeny to determine if the desired characteristic has beensuccessfully transferred.

In one embodiment, progeny lettuce plants of a backcross in which RX16883035 is the recurrent parent comprise (i) the desired trait from thenon-recurrent parent and (ii) all of the physiological and morphologicalcharacteristics of lettuce line RX 16883035 as determined at the 5%significance level when grown in the same environmental conditions.

Lettuce varieties can also be developed from more than two parents. Thetechnique, known as modified backcrossing, uses different recurrentparents during the backcrossing. Modified backcrossing may be used toreplace the original recurrent parent with a variety having certain moredesirable characteristics or multiple parents may be used to obtaindifferent desirable characteristics from each.

With the development of molecular markers associated with particulartraits, it is possible to add additional traits into an established germline, such as represented here, with the end result being substantiallythe same base germplasm with the addition of a new trait or traits.Molecular breeding, as described in Moose and Mumm, 2008 (PlantPhysiology, 147: 969-977), for example, and elsewhere, provides amechanism for integrating single or multiple traits or QTL into an eliteline. This molecular breeding-facilitated movement of a trait or traitsinto an elite line may encompass incorporation of a particular genomicfragment associated with a particular trait of interest into the eliteline by the mechanism of identification of the integrated genomicfragment with the use of flanking or associated marker assays. In theembodiment represented here, one, two, three or four genomic loci, forexample, may be integrated into an elite line via this methodology. Whenthis elite line containing the additional loci is further crossed withanother parental elite line to produce hybrid offspring, it is possibleto then incorporate at least eight separate additional loci into thehybrid. These additional loci may confer, for example, such traits as adisease resistance or a fruit quality trait. In one embodiment, eachlocus may confer a separate trait. In another embodiment, loci may needto be homozygous and exist in each parent line to confer a trait in thehybrid. In yet another embodiment, multiple loci may be combined toconfer a single robust phenotype of a desired trait.

Many single locus traits have been identified that are not regularlyselected for in the development of a new inbred but that can be improvedby backcrossing techniques. Single locus traits may or may not betransgenic; examples of these traits include, but are not limited to,male sterility, herbicide resistance, resistance to bacterial, fungal,or viral disease, insect resistance, restoration of male fertility,modified fatty acid or carbohydrate metabolism, and enhanced nutritionalquality. These comprise genes generally inherited through the nucleus.

Direct selection may be applied where the single locus acts as adominant trait. An example of a dominant trait is the downy mildewresistance trait. For this selection process, the progeny of the initialcross are sprayed with downy mildew spores prior to the backcrossing.The spraying eliminates any plants which do not have the desired downymildew resistance characteristic, and only those plants which have thedowny mildew resistance gene are used in the subsequent backcross. Thisprocess is then repeated for all additional backcross generations.

Selection of lettuce plants for breeding is not necessarily dependent onthe phenotype of a plant and instead can be based on geneticinvestigations. For example, one can utilize a suitable genetic markerwhich is closely genetically linked to a trait of interest. One of thesemarkers can be used to identify the presence or absence of a trait inthe offspring of a particular cross, and can be used in selection ofprogeny for continued breeding. This technique is commonly referred toas marker assisted selection. Any other type of genetic marker or otherassay which is able to identify the relative presence or absence of atrait of interest in a plant can also be useful for breeding purposes.Procedures for marker assisted selection applicable to the breeding oflettuce are well known in the art. Such methods will be of particularutility in the case of recessive traits and variable phenotypes, orwhere conventional assays may be more expensive, time consuming orotherwise disadvantageous. Types of genetic markers which could be usedin accordance with the invention include, but are not necessarilylimited to, Simple Sequence Length Polymorphisms (SSLPs) (Williams etal., Nucleic Acids Res., 1 8:6531-6535, 1990), Randomly AmplifiedPolymorphic DNAs (RAPDs), DNA Amplification Fingerprinting (DAF),Sequence Characterized Amplified Regions (SCARs), Arbitrary PrimedPolymerase Chain Reaction (AP-PCR), Amplified Fragment LengthPolymorphisms (AFLPs) (EP 534 858, specifically incorporated herein byreference in its entirety), and Single Nucleotide Polymorphisms (SNPs)(Wang et al., Science, 280:1077-1082, 1998).

E. Plants Derived From Lettuce Line RX 16883035 by Genetic Engineering

Many useful traits that can be introduced by backcrossing, as well asdirectly into a plant, are those which are introduced by genetictransformation techniques. Genetic transformation may therefore be usedto insert a selected transgene into the lettuce line of the invention ormay, alternatively, be used for the preparation of transgenes which canbe introduced by backcros sing. Methods for the transformation ofplants, including lettuce, are well known to those of skill in the art.

Vectors used for the transformation of lettuce cells are not limited solong as the vector can express an inserted DNA in the cells. Forexample, vectors comprising promoters for constitutive gene expressionin lettuce cells (e.g., cauliflower mosaic virus 35S promoter) andpromoters inducible by exogenous stimuli can be used. Examples ofsuitable vectors include pBI binary vector. The “lettuce cell” intowhich the vector is to be introduced includes various forms of lettucecells, such as cultured cell suspensions, protoplasts, leaf sections,and callus.

A vector can be introduced into lettuce cells by known methods, such asthe polyethylene glycol method, polycation method, electroporation,Agrobacterium-mediated transfer, particle bombardment and direct DNAuptake by protoplasts. See, e.g., Pang et al. (The Plant J., 9, 899-909,1996).

To effect transformation by electroporation, one may employ eitherfriable tissues, such as a suspension culture of cells or embryogeniccallus or alternatively one may transform immature embryos or otherorganized tissue directly. In this technique, one would partiallydegrade the cell walls of the chosen cells by exposing them topectin-degrading enzymes (pectolyases) or mechanically wound tissues ina controlled manner. An example of electroporation of lettuceprotoplasts is presented in Chupeau et al. (Bio/Tech., 7:503-508, 1989).

A particularly efficient method for delivering transforming DNA segmentsto plant cells is microprojectile bombardment. In this method, particlesare coated with nucleic acids and delivered into cells by a propellingforce. Exemplary particles include those comprised of tungsten,platinum, and preferably, gold. For the bombardment, cells in suspensionare concentrated on filters or solid culture medium. Alternatively,immature embryos or other target cells may be arranged on solid culturemedium. The cells to be bombarded are positioned at an appropriatedistance below the macroprojectile stopping plate.

An illustrative embodiment of a method for delivering DNA into plantcells by acceleration is the Biolistics Particle Delivery System, whichcan be used to propel particles coated with DNA or cells through ascreen, such as a stainless steel or Nytex screen, onto a surfacecovered with target lettuce cells. The screen disperses the particles sothat they are not delivered to the recipient cells in large aggregates.It is believed that a screen intervening between the projectileapparatus and the cells to be bombarded reduces the size of projectilesaggregate and may contribute to a higher frequency of transformation byreducing the damage inflicted on the recipient cells by projectiles thatare too large.

Microprojectile bombardment techniques are widely applicable, and may beused to transform virtually any plant species. Examples involvingmicroprojectile bombardment transformation with lettuce can be found in,for example, Elliott et al. (Plant Cell Rep., 18:707-714, 2004) andMolinier et al. (Plant Cell Rep., 21:251-256, 2002).

Agrobacterium-mediated transfer is another widely applicable system forintroducing gene loci into plant cells. An advantage of the technique isthat DNA can be introduced into whole plant tissues, thereby bypassingthe need for regeneration of an intact plant from a protoplast. ModernAgrobacterium transformation vectors are capable of replication in E.coli as well as Agrobacterium, allowing for convenient manipulations(Klee et al., Bio-Technology, 3(7):637-642, 1985). Moreover, recenttechnological advances in vectors for Agrobacterium-mediated genetransfer have improved the arrangement of genes and restriction sites inthe vectors to facilitate the construction of vectors capable ofexpressing various polypeptide coding genes. The vectors described haveconvenient multi-linker regions flanked by a promoter and apolyadenylation site for direct expression of inserted polypeptidecoding genes. Additionally, Agrobacterium containing both armed anddisarmed Ti genes can be used for transformation.

In those plant strains where Agrobacterium-mediated transformation isefficient, it is the method of choice because of the facile and definednature of the gene locus transfer. The use of Agrobacterium-mediatedplant integrating vectors to introduce DNA into plant cells is wellknown in the art (Fraley et al., Bio/Technology, 3:629-635, 1985; U.S.Pat. No. 5,563,055). For example, U.S. Pat. No. 5,349,124 describes amethod of transforming lettuce plant cells using Agrobacterium-mediatedtransformation. By inserting a chimeric gene having a DNA codingsequence encoding for the full-length B.t. toxin protein that expressesa protein toxic toward Lepidopteran larvae, this methodology resulted inlettuce having resistance to such insects.

Transformation of plant protoplasts also can be achieved using methodsbased on calcium phosphate precipitation, polyethylene glycol treatment,electroporation, and combinations of these treatments (see, e.g.,Potrykus et al., Mol. Gen. Genet., 199:183-188, 1985; Omirulleh et al.,Plant Mol. Biol., 21(3):415-428, 1993; Fromm et al., Nature,312:791-793, 1986; Uchimiya et al., Mol. Gen. Genet., 204:204, 1986;Marcotte et al., Nature, 335:454, 1988). Transformation of plants andexpression of foreign genetic elements is exemplified in Choi et al.(Plant Cell Rep., 13: 344-348, 1994) and Ellul et al. (Theor. Appl.Genet., 107:462-469, 2003).

A number of promoters have utility for plant gene expression for anygene of interest including but not limited to selectable markers,scoreable markers, genes for pest tolerance, disease resistance,nutritional enhancements and any other gene of agronomic interest.Examples of constitutive promoters useful for lettuce plant geneexpression include, but are not limited to, the cauliflower mosaic virus(CaMV) P-35S promoter, which confers constitutive, high-level expressionin most plant tissues (see, e.g., Odel et al., Nature, 313:810, 1985),including monocots (see, e.g., Dekeyser et al., Plant Cell, 2:591, 1990;Terada and Shimamoto, Mol. Gen. Genet., 220:389, 1990); a tandemlyduplicated version of the CaMV 35S promoter, the enhanced 35S promoter(P-e35S) the nopaline synthase promoter (An et al., Plant Physiol.,88:547, 1988), the octopine synthase promoter (Fromm et al., Plant Cell,1:977, 1989); and the figwort mosaic virus (P-FMV) promoter as describedin U.S. Pat. No. 5,378,619 and an enhanced version of the FMV promoter(P-eFMV) where the promoter sequence of P-FMV is duplicated in tandem,the cauliflower mosaic virus 19S promoter, a sugarcane bacilliform viruspromoter, a commelina yellow mottle virus promoter, and other plant DNAvirus promoters known to express in plant cells.

A variety of plant gene promoters that are regulated in response toenvironmental, hormonal, chemical, and/or developmental signals can beused for expression of an operably linked gene in plant cells, includingpromoters regulated by (1) heat (Callis et al., Plant Physiol., 88:965,1988), (2) light (e.g., pea rbcS-3A promoter, Kuhlemeier et al., PlantCell, 1:471, 1989; maize rbcS promoter, Schaffner and Sheen, Plant Cell,3:997, 1991; or chlorophyll a/b-binding protein promoter, Simpson etal., EMBO J., 4:2723, 1985), (3) hormones, such as abscisic acid(Marcotte et al., Plant Cell, 1:969, 1989), (4) wounding (e.g., wunl,Siebertz et al., Plant Cell, 1:961, 1989); or (5) chemicals such asmethyl jasmonate, salicylic acid, or Safener. It may also beadvantageous to employ organ-specific promoters (e.g., Roshal et al.,EMBO J., 6:1155, 1987; Schernthaner et al., EMBO J., 7:1249, 1988;Bustos et al., Plant Cell, 1:839, 1989).

Exemplary nucleic acids which may be introduced to the lettuce lines ofthis invention include, for example, DNA sequences or genes from anotherspecies, or even genes or sequences which originate with or are presentin the same species, but are incorporated into recipient cells bygenetic engineering methods rather than classical reproduction orbreeding techniques. However, the term “exogenous” is also intended torefer to genes that are not normally present in the cell beingtransformed, or perhaps simply not present in the form, structure, etc.,as found in the transforming DNA segment or gene, or genes which arenormally present and that one desires to express in a manner thatdiffers from the natural expression pattern, e.g., to over-express.Thus, the term “exogenous” gene or DNA is intended to refer to any geneor DNA segment that is introduced into a recipient cell, regardless ofwhether a similar gene may already be present in such a cell. The typeof DNA included in the exogenous DNA can include DNA which is alreadypresent in the plant cell, DNA from another plant, DNA from a differentorganism, or a DNA generated externally, such as a DNA sequencecontaining an antisense message of a gene, or a DNA sequence encoding asynthetic or modified version of a gene.

Many hundreds if not thousands of different genes are known and couldpotentially be introduced into a lettuce plant according to theinvention. Non-limiting examples of particular genes and correspondingphenotypes one may choose to introduce into a lettuce plant include oneor more genes for insect tolerance, such as a Bacillus thuringiensis(B.t.) gene, pest tolerance such as genes for fungal disease control,herbicide tolerance such as genes conferring glyphosate tolerance, andgenes for quality improvements such as yield, nutritional enhancements,environmental or stress tolerances, or any desirable changes in plantphysiology, growth, development, morphology or plant product(s). Forexample, structural genes would include any gene that confers insecttolerance including but not limited to a Bacillus insect control proteingene as described in WO 99/31248, herein incorporated by reference inits entirety, U.S. Pat. No. 5,689,052, herein incorporated by referencein its entirety, U.S. Pat. Nos. 5,500,365 and 5,880,275, hereinincorporated by reference it their entirety. In another embodiment, thestructural gene can confer tolerance to the herbicide glyphosate asconferred by genes including, but not limited to Agrobacterium strainCP4 glyphosate resistant EPSPS gene (aroA:CP4) as described in U.S. Pat.No. 5,633,435, herein incorporated by reference in its entirety, orglyphosate oxidoreductase gene (GOX) as described in U.S. Pat. No.5,463,175, herein incorporated by reference in its entirety.

Alternatively, the DNA coding sequences can affect these phenotypes byencoding a non-translatable RNA molecule that causes the targetedinhibition of expression of an endogenous gene, for example viaantisense- or cosuppression-mediated mechanisms (see, for example, Birdet al., Biotech. Gen. Engin. Rev., 9:207, 1991). The RNA could also be acatalytic RNA molecule (i.e., a ribozyme) engineered to cleave a desiredendogenous mRNA product (see for example, Gibson and Shillito, Mol.Biotech., 7:125,1997). Thus, any gene which produces a protein or mRNAwhich expresses a phenotype or morphology change of interest is usefulfor the practice of the present invention.

F. Definitions

In the description and tables herein, a number of terms are used. Inorder to provide a clear and consistent understanding of thespecification and claims, the following definitions are provided:

Allele: Any of one or more alternative forms of a gene locus, all ofwhich alleles relate to one trait or characteristic. In a diploid cellor organism, the two alleles of a given gene occupy corresponding locion a pair of homologous chromosomes.

Backcrossing: A process in which a breeder repeatedly crosses hybridprogeny, for example a first generation hybrid (F₁), back to one of theparents of the hybrid progeny. Backcrossing can be used to introduce oneor more single locus conversions from one genetic background intoanother.

Converted (Conversion) Plant: Plants which are developed by a plantbreeding technique called backcrossing, wherein essentially all of thedesired morphological and physiological characteristics of a lettuceline are recovered in addition to the trait transferred into the varietyvia the backcrossing technique and/or by genetic transformation.

Crossing: The mating of two parent plants.

Cross-pollination: Fertilization by the union of two gametes fromdifferent plants.

Diploid: A cell or organism having two sets of chromosomes.

Emasculate: The removal of plant male sex organs or the inactivation ofthe organs with a cytoplasmic or nuclear genetic factor conferring malesterility or a chemical agent.

Enzymes: Molecules which can act as catalysts in biological reactions.

F₁ Hybrid: The first generation progeny of the cross of two nonisogenicplants.

Genotype: The genetic constitution of a cell or organism.

Haploid: A cell or organism having one set of the two sets ofchromosomes in a diploid.

Linkage: A phenomenon wherein alleles on the same chromosome tend tosegregate together more often than expected by chance if theirtransmission was independent.

Marker: A readily detectable phenotype, preferably inherited incodominant fashion (both alleles at a locus in a diploid heterozygoteare readily detectable), with no environmental variance component, i.e.,heritability of 1.

Phenotype: The detectable characteristics of a cell or organism, whichcharacteristics are the manifestation of gene expression.

Quantitative Trait Loci (QTL): Quantitative trait loci (QTL) refer togenetic loci that control to some degree numerically representabletraits that are usually continuously distributed.

Regeneration: The development of a plant from tissue culture.

Royal Horticultural Society (RHS) color chart value: The RHS color chartis a standardized reference which allows accurate identification of anycolor. A color's designation on the chart describes its hue, brightnessand saturation. A color is precisely named by the RHS color chart byidentifying the group name, sheet number and letter, e.g., Yellow-OrangeGroup 19A or Red Group 41B.

Self-pollination: The transfer of pollen from the anther to the stigmaof the same plant.

Substantially Equivalent: A characteristic that, when compared, does notshow a statistically significant difference (e.g., p=0.05) from themean.

Tetraploid: A cell or organism having four sets of chromosomes.

Tissue Culture: A composition comprising isolated cells of the same or adifferent type or a collection of such cells organized into parts of aplant.

Transgene: A genetic locus comprising a sequence which has beenintroduced into the genome of a lettuce plant by transformation.

Triploid: A cell or organism having three sets of chromosomes.

G. Deposit Information

A deposit of lettuce line RX 16883035, disclosed above and recited inthe claims, has been made with the American Type Culture Collection(ATCC), 10801 University Blvd., Manassas, Va. 20110-2209. The date ofdeposit was Feb. 2, 2012. Upon issuance of a patent, all restrictionsupon the deposit will be removed, and the deposit is intended to meetall of the requirements of 37 C.F.R. §1.801-1.809. The accession numberfor those deposited seeds of lettuce line RX 16883035 is ATCC AccessionNo. PTA-12487. The deposit will be maintained in the depository for aperiod of 30 years, or 5 years after the last request, or for theeffective life of the patent, whichever is longer, and will be replacedif necessary during that period.

Although the foregoing invention has been described in some detail byway of illustration and example for purposes of clarity andunderstanding, it will be obvious that certain changes and modificationsmay be practiced within the scope of the invention, as limited only bythe scope of the appended claims.

All references cited herein are hereby expressly incorporated herein byreference.

What is claimed is:
 1. A seed of lettuce line RX 16883035, a sample ofseed of said line having been deposited under ATCC Accession NumberPTA-12487.
 2. A plant of lettuce line RX 16883035, a sample of seed ofsaid line having been deposited under ATCC Accession Number PTA-12487.3. A plant part of the plant of claim
 2. 4. The plant part of claim 3,wherein said part is selected from the group consisting of a pollen, anovule and a cell.
 5. A lettuce plant, or a part thereof, having all thephysiological and morphological characteristics of the lettuce plant ofclaim
 2. 6. A tissue culture of regenerable cells of lettuce line RX16883035, a sample of seed of said line having been deposited under ATCCAccession Number PTA-12487.
 7. The tissue culture according to claim 6,comprising cells or protoplasts from a plant part selected from thegroup consisting of embryos, meristems, cotyledons, pollen, leaves,anthers, roots, root tips, pistil, flower, seed and stalks.
 8. A lettuceplant regenerated from the tissue culture of claim 6, wherein theregenerated plant expresses all of the physiological and morphologicalcharacteristics of lettuce line RX 16883035, a sample of seed of saidline having been deposited under ATCC Accession Number PTA-12487.
 9. Amethod of producing lettuce seed, comprising crossing the plant of claim2 with a second lettuce plant.
 10. The method of claim 9, wherein theplant of lettuce line RX 16883035 is the female parent.
 11. The methodof claim 9, wherein the plant of lettuce line RX 16883035 is the maleparent.
 12. An F₁ hybrid seed produced by the method of claim
 9. 13. AnF₁ hybrid plant produced by growing the seed of claim
 12. 14. A methodfor producing a seed of a line RX 16883035-derived lettuce plantcomprising the steps of: (a) crossing a lettuce plant of line RX16883035, a sample of seed of said line having been deposited under ATCCAccession Number PTA-12487, with a second lettuce plant; and (b)allowing seed of a RX 16883035-derived lettuce plant to form.
 15. Themethod of claim 14, further comprising the steps of: (c) crossing aplant grown from said RX 16883035-derived lettuce seed with itself or asecond lettuce plant to yield additional RX 16883035-derived lettuceseed; (d) growing said additional RX 16883035-derived lettuce seed ofstep (c) to yield additional RX 16883035-derived lettuce plants; and (e)repeating the crossing and growing steps of (c) and (d) to generatefurther RX 16883035-derived lettuce plants.
 16. A method of vegetativelypropagating a plant of lettuce line RX 16883035 comprising the steps of:(a) collecting tissue capable of being propagated from a plant oflettuce line RX 16883035, a sample of seed of said line having beendeposited under ATCC Accession Number PTA-12487; and (b) producing atleast a first rooted plant from said tissue.
 17. A process of producinga conversion of lettuce line RX 16883035 comprising at least one newtrait, the process comprising: (a) crossing a plant of lettuce line RX16883035, wherein a sample of seed of said line has been deposited underATCC Accession Number PTA-12487, with a lettuce plant that comprises atleast one new trait to produce progeny seed; (b) harvesting and plantingthe progeny seed to produce at least one progeny plant of a subsequentgeneration, wherein the progeny plant comprises the at least one newtrait; (c) crossing the progeny plant with a plant of lettuce line RX16883035 to produce backcross progeny seed; (d) harvesting and plantingthe backcross progeny seed to produce at least one backcross progenyplant; and (e) repeating steps (c) and (d) for at least three additionalgenerations to produce a converted plant of lettuce line RX 16883035,wherein the converted plant of lettuce line RX 16883035 comprises the atleast one new trait.
 18. A converted lettuce plant produced by themethod of claim
 17. 19. A method of producing a plant of lettuce line RX16883035 comprising an added desired trait, the method comprisingintroducing a transgene conferring the desired trait into a plant oflettuce line RX 16883035, wherein a sample of seed of said line has beendeposited under ATCC Accession Number PTA-12487.
 20. A plant produced bythe method of claim
 19. 21. A method of producing food comprising: (a)obtaining the plant of claim 2, and (b) collecting leaf tissue from theplant, wherein the leaf tissue is capable of use as food.